In:
PLOS ONE, Public Library of Science (PLoS), Vol. 17, No. 2 ( 2022-2-25), p. e0264130-
Abstract:
The global COVID-19 pandemic has highlighted the need for rapid, accurate and accessible nucleic acid tests to enable timely identification of infected individuals. We optimized a sample-to-answer nucleic acid test for SARS-CoV-2 that provides results in 〈 1 hour using inexpensive and readily available reagents. The test workflow includes a simple lysis and viral inactivation protocol followed by direct isothermal amplification of viral RNA using RT-LAMP. The assay was validated using two different instruments, a portable isothermal fluorimeter and a standard thermocycler. Results of the RT-LAMP assay were compared to traditional RT-qPCR for nasopharyngeal swabs, nasal swabs, and saliva collected from a cohort of patients hospitalized due to COVID-19. For all three sample types, positive agreement with RT-LAMP performed using the isothermal fluorimeter was 100% for samples with Ct 〈 30 and 69–91% for samples with Ct 〈 40. Following validation, the test was successfully scaled to test the saliva of up to 400 asymptomatic individuals per day as part of the campus surveillance program at Rice University. Successful development, validation, and scaling of this sample-to-answer, extraction-free real-time RT-LAMP test for SARS-CoV-2 adds a highly adaptable tool to efforts to control the COVID-19 pandemic, and can inform test development strategies for future infectious disease threats.
Type of Medium:
Online Resource
ISSN:
1932-6203
DOI:
10.1371/journal.pone.0264130
DOI:
10.1371/journal.pone.0264130.g001
DOI:
10.1371/journal.pone.0264130.g002
DOI:
10.1371/journal.pone.0264130.g003
DOI:
10.1371/journal.pone.0264130.g004
DOI:
10.1371/journal.pone.0264130.g005
DOI:
10.1371/journal.pone.0264130.g006
DOI:
10.1371/journal.pone.0264130.g007
DOI:
10.1371/journal.pone.0264130.s001
DOI:
10.1371/journal.pone.0264130.s002
DOI:
10.1371/journal.pone.0264130.s003
DOI:
10.1371/journal.pone.0264130.s004
DOI:
10.1371/journal.pone.0264130.s005
DOI:
10.1371/journal.pone.0264130.s006
DOI:
10.1371/journal.pone.0264130.s007
DOI:
10.1371/journal.pone.0264130.s008
DOI:
10.1371/journal.pone.0264130.s009
DOI:
10.1371/journal.pone.0264130.s010
DOI:
10.1371/journal.pone.0264130.s011
DOI:
10.1371/journal.pone.0264130.s012
DOI:
10.1371/journal.pone.0264130.r001
DOI:
10.1371/journal.pone.0264130.r002
DOI:
10.1371/journal.pone.0264130.r003
DOI:
10.1371/journal.pone.0264130.r004
DOI:
10.1371/journal.pone.0264130.r005
DOI:
10.1371/journal.pone.0264130.r006
Language:
English
Publisher:
Public Library of Science (PLoS)
Publication Date:
2022
detail.hit.zdb_id:
2267670-3
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